Effect of size and processing method on the cytotoxicity of realgar nanoparticles in cancer cell lines

Weizhong Zhao¹, Xun Lu³, Yuan Yuan¹, Changsheng Liu¹, Baican Yang³, Hua Hong¹, Guoying Wang³, Fanyan Zeng^2
1The State Key Laboratory of Bioreactor Engineering, ²Key Laboratory for Ultrafine Materials of Ministry of Education and Engineering Research Center for Biomedical Materials of Ministry of Education, East China University of Science and Technology, ³Pharmacy Department of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai, People’s Republic of China

Abstract: In this study, the effects of the size and Chinese traditional processing (including elutriation, water cleaning, acid cleaning, alkali cleaning) on realgar nanoparticles (RN)-induced antitumor activity in human osteosarcoma cell lines (MG-63) and hepatoma carcinoma cell lines (HepG-2) were investigated. The human normal liver cell line (L-02) was used as control. RN was prepared by high-energy ball milling technology. The results showed that with the assistance of sodium dodecyl sulfate, the size of realgar could be reduced to 127 nm after 12 hours’ ball milling. The surface charge was decreased from 0.83 eV to −17.85 eV and the content of As₂O₃ clearly increased. Except for elutriation, the processing methods did not clearly change the size of the RN, but the content of As₂O₃ was reduced dramatically. In vitro MTT tests indicated that in the two cancer cell lines, RN cytotoxicity was more intense than that of the coarse realgar nanoparticles, and cytotoxicity was typically time- and concentration-dependent. Also, RN cytotoxicities in the HepG-2 and L-02 cells all increased with increasing milling time. Due to the reduction of the As₂O₃ content, water cleaning, acid cleaning, and alkali cleaning decreased RN cytotoxicity in HepG-2, but RN after elutriation, with the lowest As₂O₃ (3.5 mg/g) and the smallest size (109.3 nm), showed comparable cytotoxicity in HepG-2 to RN without treatment. Meanwhile, RN-induced cytotoxicity in L-02 cells was clearly reduced. Therefore, it can be concluded that RN may provide a strong antiproliferation effect in the MG-63 and HepG-2 cells. Elutriation processing is a suitable approach to limit the dangerous side-effects of As₂O₃, while maintaining the effectiveness of RN.

Keywords: realgar nanoparticles, cytotoxicity, size, processing, apoptosis